Investigation of the effects and mechanisms of sucralfate on the intestinal epithelium in an experimental in-vitro model of necrotizing enterocolitis

Objective: To investigate the effects and mechanisms of sucralfate on enterocytes in an experimental in-vitro model of necrotizing enterocolitis (NEC). Materials and Methods: 5 experimental groups were formed by using intestinal epithelial cell line (IEC-6). NEC induction was performed with lipopolysaccharide (LPS) and. TNF-α and IL-8 levels were measured by ELISA. The non-toxic dose of sucralfate was determined by the MTT method. The control group (KG) consisted of IEC-6 only. IEC-6 were incubated with sucralfate in the sham group (SG) and with LPS in the NEC group for 48 hours. In the treatment group (TG), after 48 hours of LPS, 48 hours of sucralfate; in the prophylaxis group (PG) after 48 hours of sucralfate, 48 hours of LPS was applied. TUNEL staining was performed on the samples. H scores of Caspase-3-8 and 9, RIPK1, RIPK3, MLKL, occludin, claudin, ICAM-1 and MadCAM-1 were calculated. Differences between groups were compared with the one-way-ANOVA test. P<0.05 values were considered significant. Results: TNF-α and IL-8 levels were higher in the NEC group compared to CG (p<0.05). The application dose of sucralfate was 106μL/cm2 at 2:1 dilution. While TUNEL positive cells were 65.6±8.26% in NEC group, 15.4±3.28% in TG and 23.4±4.21 in PG (p<0.05). There was no significant difference between the H scores of Caspase3. Caspase 8-9 and RIPK1 were elevated only in the NEC group (p<0.05). There was a decrease in RIPK3 when the NEK group was compared with both TG and PG (p<0.05). MLKL expression was higher in NEC group compared to KG (p<0.05) and lower than TG and PG (p<0.05). ICAM-1 expression did not differ significantly between the groups. MadCAM-1 was higher in NEC group compared to KG (p<0.05), sucralfate administration decreased MadCAM-1 expression in both PG and TG (p<0.05). Occludin distribution was higher in TG, while claudin distribution was lower (p<0.05). Conclusion: In in-vitro NEC model, apoptosis and necroptosis increase and the expression of cell adhesion molecules changes. Sucralfate administration suppresses apoptosis and necroptosis and helps protect cell adhesion molecules. Prophylactic administration of sucralfate does not appear to be as effective as therapeutic administration.